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Melting or thermal denaturation of a DNA molecule and the different bubble-rich, premelting DNA states that serve as a precursor for DNA thermal denaturation are vital events in DNA thermophysics. In this study, we employ cantilever-based sensing to firstly pinpoint the occurrence of DNA melting and identify the temperature Tm characterizing the melting. Very importantly, this sensing is carried out with an extremely small volume (∼picoliters) of DNA sample with the cantilever demonstrating an extremely high sensitivity on the order of mJ / g ⋅ K corresponding to pico-Joules of energy input. Secondly, this same large sensitivity of the cantilever is used to quantify the hitherto unknown thermophysical properties of the bubble-rich DNA premelting states. In fact, for both the melting and premelting states, the cantilever provides a framework to calculate the specific heat capacity and the storage and loss moduli of the cantilever-DNA-solution system, thereby establishing a platform for quantifying DNAs' thermo-mechanical behavior. [ABSTRACT FROM AUTHOR]

Growth factor receptor-bound protein 2 (Grb2) have been proved by a lot of studies playing a major role in cell proliferation and cell differentiation. However, the regulation of Grb2 expression by microRNAs (miRNAs) in chicken breast muscle still remains unknown. The expression profile of Grb2 was checked based on our previous RNA sequencing data and the Grb2 relative expression level in breast muscle of aged hens (55-week-old) was validated significantly higher than juvenile hens (20-week-old) using qRT-PCR. miRNAs that interact with Grb2 have been predicted in chicken and the relationship between the potential miRNA and Grb2 was verified using dual luciferase reporter assay in chicken DF1 cells. Dual-luciferase reporter assays results demonstrated that the expression of luciferase reporter gene linked with part sequence of the 3′UTR of chicken Grb2 gene was down-regulated by the overexpression of gga (Gallus Gallus)-miR-200a-3p in the DF1 cells, and the down-regulation behavior was abolished when the gga-miR-200a-3p binding site in 3′UTR of Grb2 was mutated, indicating that gga-miR-200a can suppress the expression level of its target gene Grb2. Therefore, we concluded that the significantly increased expression level of Grb2 in the breast muscle of aged chicken can (at least partly can) be explained by the decreased expression of miR-200a, which reduced the inhibitory effect on Grb2. Taken together, these findings suggest that gga-miR-200a can suppress the expression level of its target gene Grb2 and might be involved in the cell differentiation and proliferation of chicken breast muscle through binding with the 3’UTR of Grb2. [ABSTRACT FROM AUTHOR]

We report a simple, rapid and cost effective detection technique for Escherichia coli ( E. coli ) using a Light Addressable Potentiometric Sensor integrated with electrospun poly acrylic acid/polyvinyl alcohol (PAA/PVA) hydrogel nanofibers as a sensing layer (NF-LAPS). Changes in pH of the media are detected as E. coli cells ferment sugar molecules and increase acidity of the surrounding. A super-Nernstian response of a 74 mV/pH change in the NF-LAPS provides high sensitivity towards E. coli with a theoretical limit of detection (LOD) of 20 CFU/ml. The high sensitivity is associated with the pH sensitive behavior of the NFs on the surface. Detection of acidic species released in cellular metabolism of pathogens in the presence of carbon source renders the sensing mechanism cheaper, less time consuming and more practical than the antibody-antigen interaction based approach. Selectivity towards E. coli for applications in drinking water detection is ensured by the incorporation of d -mannose for specific binding. [ABSTRACT FROM AUTHOR]

Ligand-directed targeting and capturing of cancer cells is a new approach for detecting circulating tumor cells (CTCs). Ligands such as antibodies have been successfully used for capturing cancer cells and an antibody based system (CellSearch®) is currently used clinically to enumerate CTCs. Here we report the use of a peptide moiety in conjunction with a microcantilever array system to selectively detect CTCs resulting from cancer, specifically breast cancer. A sensing microcantilever, functionalized with a breast cancer specific peptide 18-4 (WxEAAYQrFL), showed significant deflection on cancer cell (MCF7 and MDA-MB-231) binding compared to when exposed to noncancerous (MCF10A and HUVEC) cells. The peptide-functionalized microcantilever allowed efficient capture and detection of cancer cells in MCF7 spiked human blood samples emulating CTCs in human blood. A detection limit of 50-100 cancer cells mL-1 from blood samples was achieved with a capture yield of 80% from spiked whole blood samples. The results emphasize the potential of peptide 18-4 as a novel peptide for capturing and detecting cancer cells in conjunction with nanomechanical cantilever platform. The reported peptide-based cantilever platform represents a new analytical approach that can lead to an alternative to the various detection platforms and can be leveraged to further study CTCs. [ABSTRACT FROM AUTHOR]

Broad-spectrum biosensing technologiesexamine sensor signals usingbiomarkers, such as proteins, DNA, antibodies, specific cells, andmacromolecules, based on direct- or indirect-conformational changes.Here, we have investigated the pH-dependent conformational isomerizationof human serum albumin (HSA) using microcantilevers as a sensing platform.Native and denatured proteins were immobilized on cantilever surfacesto understand the effect of pH on conformational changes of the proteinwith respect to the coupling ligand. Our results show that protonationand deprotonation of amino acid residues on proteins play a significantrole in generating charge-induced cantilever deflection. Surface plasmonresonance (SPR) was employed as a complementary technique to validatethe results. [ABSTRACT FROM AUTHOR]

Real-time, label-free detection of Gram-positive bacteria with high selectivity and sensitivity is demonstrated using an interdigitated impedimetric array functionalized with naturally produced antimicrobial peptide from class IIa bacteriocins. The antimicrobial peptide, leucocin A, was chemically synthesized and covalently immobilized on interdigitated gold microelectrodes via the interaction between the C-terminal carboxylic acid of the peptide and free amines of a preattached thiolated linker. Exposing the peptide sensor to various concentrations of Gram-positive bacteria generated reproducible impedance spectra that detected peptide–bacteria interactions at a concentration of 1 cell/μL. The peptide sensor also selectively detected Listeria monocytogenes from other Gram-positive strains at a concentration of 103 cfu mL–1. The study highlights that short peptide ligands from bacteriocin class offer high selectivity in bacterial detection and can be used in developing a robust, portable biosensor device to efficiently detect pathogenic Gram-positive bacteria in food samples. [ABSTRACT FROM AUTHOR]

Egg-laying performance is one of the most important economic traits in the poultry industry. Commercial layers can lay one egg almost every day during their peak-laying period. However, many Chinese indigenous chicken breeds show a relatively low egg-laying rate, even during their peak-laying period. To understand what makes the difference in egg production, we compared the hypothalamus transcriptome profiles of Lushi blue-shelled-egg chickens (LBS), a Chinese indigenous breed with low egg-laying rate and Rhode Island Red chickens (RIR), a commercial layer with relatively high egg-laying rate using RNA-seq. A total of 753 differentially expressed genes (DEGs) were obtained. Of these DEGs, 38 genes were enriched in 2 Gene Ontology (GO) terms, namely reproduction term and the reproductive process term, and 6 KEGG pathways, namely Wnt signaling pathway, Oocyte meiosis, GnRH signaling pathway, Thyroid hormone signaling pathway, Thyroid hormone synthesis and MAPK signaling pathway, which have been long known to be involved in egg production regulation. To further determine the core genes from the 38 DEGs, protein-protein interaction (PPI) network, co-expression network and transcriptional regulatory network analyses were carried out. After integrated analysis and experimental validation, 4 core genes including RAC1, MRE11A, MAP7 and SOX5 were identified as the potential core genes that are responsible for the laying-rate difference between the 2 breeds. These findings paved the way for future investigating the mechanism of egg-laying regulation and enriched the chicken reproductive regulation theory. [ABSTRACT FROM AUTHOR]

Electrocatalytic nitrogen (N2) reduction reaction (NRR) presents a sustainable alternative to the Haber–Bosch process for ammonia (NH3) synthesis. Iron phthalocyanine (FePc) is demonstrated as a promising catalyst for the electrocatalytic NRR. However, FePc with planar symmetric Fe‐N4 sites exhibits poor N2 adsorption and activation capabilities, resulting in an unsatisfactory NRR performance. Herein, an axial oxygen coordination strategy is developed to optimize the local electron distribution on FePc for improving N2 adsorption and activation. The as‐obtained FePc‐O‐CP shows a superior NH3 yield rate (59.72 µg h−1 mg−1cat.) and a considerable Faradaic efficiency (13.76%) in 0.1 m HCl. Density functional theory (DFT) calculations verify that the axial oxygen ligand on FePc inhibits the adsorption of H+ and enhances the N2 adsorption and activation, thereby greatly promoting NH3 generation. This work reveals the significance of regulating the local coordination environment of single‐atom catalysts for improving electrocatalytic NRR performance and provides a feasible strategy for the rational design of atomic‐scale active sites. [ABSTRACT FROM AUTHOR]

Background: Estrogen plays an essential role in female development and reproductive function. In chickens, estrogen is critical for lipid metabolism in the liver. The regulatory molecular network of estrogen in chicken liver is poorly understood. To identify estrogen-responsive genes and estrogen functional sites on a genome-wide scale, we determined expression profiles of mRNAs, lncRNAs, and miRNAs in estrogen-treated ((17β-estradiol)) and control chicken livers using RNA-Sequencing (RNA-Seq) and studied the estrogen receptor α binding sites by ChIP-Sequencing (ChIP-Seq). Results: We identified a total of 990 estrogen-responsive genes, including 962 protein-coding genes, 11 miRNAs, and 17 lncRNAs. Functional enrichment analyses showed that the estrogen-responsive genes were highly enriched in lipid metabolism and biological processes. Integrated analysis of the data of RNA-Seq and ChIP-Seq, identified 191 genes directly targeted by estrogen, including 185 protein-coding genes, 4 miRNAs, and 2 lncRNAs. In vivo and in vitro experiments showed that estrogen decreased the mRNA expression of PPARGC1B, which had been reported to be linked with lipid metabolism, by directly increasing the expression of miR-144-3p. Conclusions: These results increase our understanding of the functional network of estrogen in chicken liver and also reveal aspects of the molecular mechanism of estrogen-related lipid metabolism. [ABSTRACT FROM AUTHOR]

Long noncoding RNAs (lncRNAs) play important roles in transcriptional and posttranscriptional regulation. However, the effects of lncRNAs on the meat quality of chicken hasn’t been elucidated clearly yet. Gushi chickens are popular in China because of their superior meat quality, particularly the tender flesh, and unique flavor. Gushi chickens are popular in China because of their superior meat quality, delicate flesh, and unique flavor. We performed RNA-Seq analysis of breast muscle from Gushi chicken at two physiological stages, including juvenile (G20W) and laying (G55W). In total, 186 lncRNAs and 881 mRNAs were differentially expressed between G20W and G55W (fold change ≥ 2.0, P < 0.05). Among them, 131 lncRNAs presented upregulated and 55 were downregulated. We identified the cis and trans target genes of the differentially expressed lncRNAs, and constructed lncRNA-mRNA interaction networks. The results showed that differentially expressed mRNAs and lncRNAs were mainly involved in ECM-receptor interaction, glycerophospholipid metabolism, ubiquitin-mediated proteolysis, and the biosynthesis of amino acids. In summary, our study utilized RNA-seq analysis to predict the functions of lncRNA on chicken meat quality. Furthermore, comprehensive analysis identified lncRNAs and their target genes, which may contribute to a better understanding of the molecular mechanisms underlying in poultry meat quality and provide a theoretical basis for further research. [ABSTRACT FROM AUTHOR]

Spontaneous self-assemblies of biomolecules can generate geometrical patterns. Our findings provide an insight into the mechanism of self-assembled ring pattern generation by human serum albumin (HSA). The self-assembly is a process guided by kinetic and thermodynamic parameters. The generated protein ring patterns display a behavior which is geometrically related to a n-simplex model and is explained through thermodynamics and chemical kinetics. [ABSTRACT FROM AUTHOR]

[Display omitted] • Sn layers effectively uniformize Zn deposition and inhibit side reactions. • Flame normalizing enriches Sn (002) facet which possesses high Zn affinity and surface wettability. • Flame normalizing strengthens the interaction between Sn layers and Zn electrodes. Flame normalizing is innovatively applied to enrich the preferential crystal plane of Zn-affinitive Sn layer on Zn anode and enhance Sn/Zn interaction concurrently, thus resulting in robust and stable Zn metal anodes. Theoretical calculations and experimental results reveal that such Sn layers effectively inhibit parasitic reactions and dendrite growth, lower Zn nucleation/deposition energy barriers, enable sufficient contact with electrolytes, and have fast reaction kinetics. Interestingly, we find that flame normalizing treatment increases Sn (200) facet which has high surface energy, thus further improving surface wettability and homogenizing Zn2+ flux. In addition, the strengthened Sn/Zn interaction enhances the structural integrity of electrodes to withstand harsh conditions. These features ensure flame normalized Sn layer covered Zn anode to deliver low Zn plating/stripping overpotential and polarization, together with a long cycling lifetime over 1200 h. The resulting Zn-based anode also significantly improves the performance of Zn//MnO 2 full cells. Overall, this work provides insights for crystal plane engineering towards stable Zn anodes. [ABSTRACT FROM AUTHOR]

Despite the advantages of the fiber‐shaped Zn‐ion microbattery (FZMB) in powering wearable electronics, several fundamental challenges hinder its practical application, mainly including dendrite growth on Zn anodes (leading to short cycle life) and low electrical conductivity of cathode (resulting in poor rate performance). Herein, a facile approach of sputtering a nano‐thin conductive carbon layer on Zn anode to effectively suppress dendrite growth and a dual‐conductive polymer strategy to fabricate ultraconductive core‐sheath fiber cathode (poly(3,4‐ethylenedioxythiophene)‐poly(styrene sulfonate) (PEDOT:PSS) fiber@polyaniline nanobulges) are demonstrated. The carbon layer suppresses Zn dendrites by uniformizing surface electric field and providing abundant nucleation sites. The superior conductivity of the cathode is inherited from two conductive polymers (in particular, PEDOT:PSS fibers have an ultrahigh conductivity of 3676 S cm−1) and their strong intermolecular interactions. The resulting FZMB shows excellent stability (over 100% capacity retention after 3000 cycles) and supercapacitor‐level rate performance (73% capacity retention from 0.1 to 10 A g−1). Kinetics and mechanism studies reveal that the surface‐controlled dual‐ion migration mechanism is also correlated with the high rate performance. The corresponding quasi‐solid‐state device exhibits high stability under extreme deformation conditions and superior water‐proof capability (94.6% capacity retention after 12 h underwater immersion), demonstrating great practical application potential. [ABSTRACT FROM AUTHOR]

[Display omitted] • The functions of reactive sites in electrocatalytic NRR and the electrocatalytic NRR mechanism are introduced. • The typical strategies to construct and characterize atomic-level reactive sites are summarized. • The recent theoretical and experimental advances of single-atoms, dual-atoms, metal clusters, vacancies, and dopants in electrocatalytic NRR are discussed. • The rigorous protocols for electrocatalytic NRR are listed. • The challenges and perspectives of atomic-level reactive sites in electrocatalytic NRR are proposed. Ammonia (NH 3) is an integral part to modern agriculture and industry. As an economical and sustainable process to convert nitrogen to NH 3 under ambient conditions, the electrocatalytic nitrogen reduction reaction (NRR) strategy has attracted considerable attention in recent years. The fabrication of atomic-level reactive sites provides an opportunity to develop novel atomic-scale catalysts with excellent electrocatalytic NRR performance. In particular, the design of atomic-level reactive sites can not only improve the activity, selectivity, and durability for electrocatalytic NRR, but also deepen the understanding of the reaction mechanism. In this review, the roles of reactive sites in electrocatalytic NRR and the electrocatalytic NRR mechanism are introduced first. Then the typical strategies to construct and characterize atomic-level reactive sites are summarized. Next, the recent progress in rational design and development of atomic-level reactive sites for electrocatalytic NRR is summarized and discussed, with a focus on single-atoms, dual-atoms, metal clusters, vacancies, and dopants. Additionally, the rigorous protocols for electrocatalytic NRR are listed. Finally, the challenges and perspectives of atomic-level reactive sites in electrocatalytic NRR are proposed to develop more credible high-efficiency NRR electrocatalysts. [ABSTRACT FROM AUTHOR]

The elongation of very long chain fatty acids protein 6 (ELOVL6) encodes a fatty acid elongase that is responsible for the final step in endogenous saturated fatty acid synthesis and involves in de novo lipogenesis. Though the regulatory mechanism of ELOVL6 expression has been studied extensively, little is known about the role of miRNA in regulating ELOVL6 gene expression in chicken until now. To investigate the regulatory mechanism of miRNA on the expression of ELOVL6 gene, bioinformatics analysis was employed to predict the potential miRNAs that binding with the 3′untranslated region (3′UTR) of ELOVL6. The putative miRNA was further screened by comparative analysis with previous miRNA-seq results. Gga-miR-22-3p, which could bind with the 3′UTR of ELOVL6 and showed negative expression correlation with ELOVL6 gene in chicken liver, was obtained. Tissue expression profiles showed that gga-miR-22-3p and ELOVL6 are extensively expressed in many tissues, and ELOVL6 with high expression level in kidney and liver tissues, and gga-miR-22-3p with high expression in lung and heart. Dual-luciferase reporter assays results indicated that the expression of luciferase reporter gene linked with part sequence of the 3′UTR of chicken ELOVL6 gene was down-regulated by the overexpression of gga-miR-22-3p in the DF1 cells, and the down-regulation behavior was abolished when the gga-miR-22-3p binding site in 3′UTR of ELOVL6 was mutated ( P > 0.05). Furthermore, the ELOVL6 expression in chicken hepatocytes was down-regulated when miR-22-3p was over-expressed. Therefore, we concluded that miR-22-3p might involve in controlling the hepatic lipid composition through affecting the expression of ELOVL6 gene, and could serve as a regulator of lipid metabolism in the liver of egg-laying hen. [ABSTRACT FROM AUTHOR]

Apolipoprotein B (ApoB) is a major protein component of plasma lipoproteins. It is involved in many important biological processes such as lipid transportation, enzyme activity regulation, and receptor recognition. Extensive studies have shown that the expression of ApoB is regulated at multiple levels. However, the regulation of ApoB expression by microRNAs (miRNAs) still remains unknown. In the present study, identified are miRNAs that are predicted to interact with ApoB in chicken. The predicted relationship between the identified miRNAs and ApoB was verified through dual luciferase reporter assay in chicken DF1 cells, and the effect of miRNAs on ApoB expression was analyzed in chicken embryo hepatocytes stimulated by 17β-estradiol. The results show that miR-101-2-5p was predicted to interact with ApoB. Dual luciferase reporter assay together with the miR-101-2-5p mimics study demostrate that ApoB is the target of miR-101-2-5p, which suppresses the expression of ApoB through binding with the 3′UTR of ApoB. Our experiments suggest that miR-101-2-5p might be involved in lipid metabolism through binding to the 3′UTR of ApoB in the liver of egg-laying chickens. [ABSTRACT FROM AUTHOR]

The cutting forces are studied for the reaming of casting aluminium alloy ZL102 (ZAlSi12) using a four-flute PCD reamer on DMG DMU 50 linear five-axis CNC machine. Kistler Dynamometer is used to measure the cutting force ( X-, Y- and Z-axes) and torque. Three-coordinate measuring machine is used for measuring cylindricity of machined hole. The cutting force signatures are diagnosed by using Fast Fourier Transform (FFT) algorithm. FFT analysis suggests that fundamental frequency is only determined by cutting speed. The frequency for the thrust signal of cutting force is zero which illustrates that the thrust is constant when cutting speed and cutting feed are fixed. The relationship between the cutting force and cutting parameter are investigated by using FFT filtering. High value of cutting feed and the low cutting speed provide the higher thrust. The optimized machining parameters are provided relative to cutting force ( Fx and Fy) and the cylindricity of reamed holes. [ABSTRACT FROM AUTHOR]

• Chicken ACSL2 was evolutionally lost. • Chicken ACSLs were evolutionally clustered into ACSL1/5/6 and ACSL3/4. • Decreased ACSL1 and increased ACSL5 expression levels were detected in the liver of peak- versus pre-laying hens. • Estrogen could transcriptionally inhibit ACSL1/3/4 and activate ACSL6. Long chain acyl-CoA synthetases (ACSLs), which drive the conversion of long chain fatty acid into acyl-CoA, an ingredient of lipid synthesis, have been well-acknowledged to exert an indispensable role in many metabolic processes in mammals, especially lipid metabolism. However, in chicken, the evolutionary characteristics, expression profiles and regulatory mechanisms of ACSL gene family are rarely understood. Here, we analyzed the genomic synteny, gene structure, evolutionary event and functional domains of the ACSL gene family members using bioinformatics methods. The spatiotemporal expression profiles of ACSL gene family, and their regulatory mechanism were investigated via bioinformatics analysis incorporated with in vivo and in vitro estrogen-treated experiments. Our results indicated that ACSL2 gene was indeed evolutionarily lost in the genome of chicken. Chicken ACSLs shared an AMP-binding functional domain, as well as highly conversed ATP/AMP and FACS signature motifs, and were clustered into two clades, ACSL1/5/6 and ACSL3/4, based on high sequence similarity, similar gene features and conversed motifs. Chicken ACSLs showed differential tissue expression distributions, wherein the significantly decreased expression level of ACSL1 and the significantly increased expression level of ACSL5 were found, respectively, the expression levels of the other ACSL members remained unchanged in the liver of peak-laying hens versus pre-laying hens. Moreover, the transcription activity of ACSL1 , ACSL3 and ACSL4 was silenced and ACSL6 was activated by estrogen, but no response to ACSL5. In conclusion, though having highly conversed functional domains, chicken ACSL gene family is organized into two separate groups, ACSL1/5/6 and ACSL3/4 , and exhibits varying expression profiles and estrogen effects. These results not only pave the way for better understanding the specific functions of ACSL genes in avian lipid metabolism, but also provide a valuable evidence for gene family characteristics. [ABSTRACT FROM AUTHOR]

Mechanical signaling involved in molecular interactions lies at the heart of materials science and biological systems, but the mechanisms involved are poorly understood. Here we use nanomechanical sensors and intact human cells to provide unique insights into the signaling pathways of connectivity networks, which deliver the ability to probe cells to produce biologically relevant, quantifiable and reproducible signals. We quantify the mechanical signals from malignant cancer cells, with 10 cells per ml in 1000-fold excess of non-neoplastic human epithelial cells. Moreover, we demonstrate that a direct link between cells and molecules creates a continuous connectivity which acts like a percolating network to propagate mechanical forces over both short and long length-scales. The findings provide mechanistic insights into how cancer cells interact with one another and with their microenvironments, enabling them to invade the surrounding tissues. Further, with this system it is possible to understand how cancer clusters are able to co-ordinate their migration through narrow blood capillaries. Samadhan Patil et al. report a new method for improving the sensitivity and reproducibility of mechanobiological measurements in malignant cancer cells. Their findings provide insight into the interaction of cells with each other and the microenvironment and may impact our understanding of metastasis. [ABSTRACT FROM AUTHOR]